Bioprinting Cell-Laden Hydrogels for Studies of Epithelial Tissue Morphogenesis

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 2805; p. 113
• Main Authors: Nerger, Bryan A, Nelson, Celeste M
• Format: Journal Article
• Language: English
• Published: United States 2024
• Subjects: Animals; Bioprinting - methods; Collagen - chemistry; Collagen - metabolism; Dogs; Drug Combinations; Epithelial Cells - cytology; Epithelial Cells - metabolism; Epithelium - growth & development; Epithelium - metabolism; Extracellular Matrix - chemistry; Extracellular Matrix - metabolism; Humans; Hydrogels - chemistry; Laminin - chemistry; Morphogenesis; Printing, Three-Dimensional; Proteoglycans - chemistry; Proteoglycans - metabolism; Tissue Engineering - methods; Tissue Scaffolds - chemistry; Biomaterial; 3D printing; Flow-induced alignment
• ISSN: 1940-6029
• DOI: 10.1007/978-1-0716-3854-5_7

The extracellular matrix (ECM) provides dynamic structural and molecular signals that affect the form and function of developing tissues. In order to parse how the individual features of the ECM impact cell- and tissue-level behavior during development, engineered culture models should reproduce key structural and molecular features of native ECM. Here, we describe a protocol for bioprinting epithelial cell aggregates embedded within a collagen-Matrigel ink in order to study the dynamic interplay between epithelial tissues and aligned networks of type I collagen fibers. Collagen fiber alignment and geometry can be spatially controlled by modulating the printing speed, nozzle geometry, surface chemistry, and degree of molecular crowding in the printing ink. We provide detailed procedures for generating epithelial cell aggregates, microextrusion printing collagen-Matrigel bioinks, culturing the three-dimensional (3D)-printed tissues, and imaging 3D-printed collagen-Matrigel constructs.